DNA methylation pattern of CALCA in preterm neonates with bacterial sepsis as a putative epigenetic biomarker

Diagnosis of bacterial sepsis in preterm neonates can be difficult when using serum markers that rely on physiological changes because these changes may not necessarily be the result of bacterial infections alone. This retrospective investigation explores the potential use of the DNA methylation pattern of CpG sites in the promoter region of the calcitonin-related polypeptide α (CALCA) gene as an epigenetic biomarker for bacterial sepsis in preterm newborns. Four novel changes in the DNA methylation of eight CpG sites were detected in this gene and are present only in neonates with bacterial sepsis: (1) partial methylation at -769 CpG in gram-negative or gram-positive early onset sepsis (EOS) and late onset sepsis (LOS) episodes; (2) demethylation of 8 CpGs in gram-negative EOS followed by LOS (ELS) and in gram-negative EOS; (3) demethylation of 7 CpGs in gram-positive ELS and gram-positive EOS; (4) -771 C:G > T:A; 5′ de novo -778 CpG mutation on both alleles in EOS. These changes were not detected in birth weight and gestational age matched controls or in newborns with isolated infections. Our results indicate that the DNA methylation pattern of the promoter region of the CALCA gene varies in different types of bacterial preterm sepsis, thus suggesting a potential use as an epigenetic biomarker. A prospective confirmation of these results is essential.     

Autofluorescence excitation‐emission matrices as a quantitative tool for the assessment of meat quality

Commercially produced meat is currently graded by a complex and partly subjective multiparameter methodology; a quantitative method of grading, using small samples would be desirable. Here, we investigate the correlation between commercial grades of beef and spectral signatures of native fluorophores in such small samples. Beef samples of different commercial grades were characterized by fluorescence spectroscopy complemented by biochemical and histological assessment. The excitation‐emission matrices of the specimens reveal five prominent native autofluorescence signatures in the excitation range from 250 to 350 nm, derived mainly from tryptophan and intramuscular fat. We found that these signatures reflect meat grade and can be used for its determination.      

Common Effects on Cancer Cells Exerted by a Random Positioning Machine and a 2D Clinostat

In this study we focused on gravity-sensitive proteins of two human thyroid cancer cell lines (ML-1; RO82-W-1), which were exposed to a 2D clinostat (CLINO), a random positioning machine (RPM) and to normal 1g-conditions. After a three (3d)- or seven-day-culture (7d) on the two devices, we found both cell types growing three-dimensionally within multicellular spheroids (MCS) and also cells remaining adherent (AD) to the culture flask, while 1g-control cultures only formed adherent monolayers, unless the bottom of the culture dish was covered by agarose. In this case, the cytokines IL-6 and IL-8 facilitated the formation of MCS in both cell lines using the liquid-overlay technique at 1g. ML-1 cells grown on the RPM or the CLINO released amounts of IL-6 and MCP-1 into the supernatant, which were significantly elevated as compared to 1g-controls. Release of IL-4, IL-7, IL-8, IL-17, eotaxin-1 and VEGF increased time-dependently, but was not significantly influenced by the gravity conditions. After 3d on the RPM or the CLINO, an accumulation of F-actin around the cellular membrane was detectable in AD cells of both cell lines. IL-6 and IL-8 stimulation of ML-1 cells for 3d and 7d influenced the protein contents of ß₁-integrin, talin-1, Ki-67, and beta-actin dose-dependently in adherent cells. The ß₁-integrin content was significantly decreased in AD and MCS samples compared with 1g, while talin-1 was higher expressed in MCS than AD populations. The proliferation marker Ki-67 was elevated in AD samples compared with 1g and MCS samples. The ß-actin content of R082-W-1 cells remained unchanged. ML-1 cells exhibited no change in ß-actin in RPM cultures, but a reduction in CLINO samples. Thus, we concluded that simulated microgravity influences the release of cytokines in follicular thyroid cancer cells, and the production of ß₁-integrin and talin-1 and predicts an identical effect under real microgravity conditions.     

Somatic embryogenesis in strawberry (<Emphasis Type="Italic">Fragaria</Emphasis> sp.) through callus culture

Somatic embryogenesis was achieved from callus from leaf and nodal segments of strawberry clone pbgel-2000. The highest percent of cultures with somatic embryos was achieved on MS medium supplemented with 1.0 mg/l 2,4-D, 0.5 mg/l BAP and 50% proline. Darkness was the best condition for incubation with daily light periods over 6 h reducing the frequency of embryogenesis. Regenerated plants (90–95%) were successfully transferred to soil, showed normal morphology and produced fruit four months after planting.     

Human–Tiger Conflict in Context: Risks to Lives and Livelihoods in the Bangladesh Sundarbans

People’s perceptions of the risk posed by wild animals to human lives and/or livelihoods can influence the rate at which people intentionally kill these species. Consequently, human–wildlife conflict (HWC) management strategies may benefit from the inclusion of actions which reduce risk perceptions. This study uses Participatory Risk Mapping (PRM) and semi-structured interviews to explore local perceptions and the wider socio-economic context of human–tiger conflict (HTC) in the Bangladesh Sundarbans area. Of the 24 locally-relevant problems identified by the PRM process, tigers were the only problem to be cited by >50 % of respondents. The ‘tiger problem’ was also perceived by villagers to be of relatively high severity. Negative perceptions of tigers in the Sundarbans communities are exacerbated by other locally-experienced poverty-related problems, as well cyclones, floods and soil erosion. Interactions between the problems experienced by villagers, including HTC, result in a complex ‘risk web’ which detrimentally affects lives and livelihoods and ultimately perpetuates poverty levels in the Sundarbans communities. This research demonstrates that PRM and in-depth, qualitative research can enhance understanding of the perceived magnitude and wider socio-economic context of risks from wildlife and aid the identification of risk perception management actions which may help to reduce the number of animals killed by people.     

Accumulation of arsenic in soil and rice under wetland condition in Bangladesh

Shallow tube well (STW) water, often contaminated with arsenic (As), is used extensively in Bangladesh for irrigating rice fields in the dry season, leading to potential As accumulation in soils. In the current study the consequences of arsenic from irrigation water and direct surface (0–15 cm) soil application were studied under field conditions with wetland rice culture over 2 years. Twenty PVC cylinders (30-cm length and 30-cm diameter) were installed in field plots to evaluate the mobility and vertical distribution of soil As, As mass balance, and the resulting influences on rice yield and plant-As concentration in Boro (dry season) and transplanted (T.) Aman (wet season) rice over the 2-year growth cycle. Treatments included irrigation-water As concentrations of 0, 1 and 2 mg L^?1 (Boro season only) and soil-As concentrations of 10 and 20 mg kg^?1. Following the 2-year cropping sequence the major portion (39.3–47.6%) of the applied arsenic was retained within the rooting zone at 0–15 cm depth, with 14.7–19.5% of the total applied As at the 5–10 cm and 10–15 cm soil depths compared to 1.3–3.6% at the 35–40 cm soil depth. These results indicate the relatively low mobility of applied As and the likely continued detrimental accumulation of As within the rooting zone. Arsenic addition in either irrigation water or as soil-applied As resulted in yield reductions from 21 to 74 % in Boro rice and 8 to 80 % in T. Aman rice, the latter indicating the strong residual effect of As on subsequent crops. The As concentrations in rice grain (0.22 to 0.81 µg g^?1), straw (2.64 to 12.52 µg g^?1) and husk (1.20 to 2.48 µg g^?1) increased with increasing addition of As. These results indicate the detrimental impacts of continued long-term irrigation with As-contaminated water on agricultural sustainability, food security and food quality in Bangladesh. A critical need exists for the development of crop and water management strategies to minimize potential As hazard in wetland rice production.